This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Norman, A.
Right arrow Articles by Sørensen, S. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Norman, A.
Right arrow Articles by Sørensen, S. J.
Agricola
Right arrow Articles by Norman, A.
Right arrow Articles by Sørensen, S. J.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, May 2005, p. 2338-2346, Vol. 71, No. 5
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.5.2338-2346.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Construction of a ColD cda Promoter-Based SOS-Green Fluorescent Protein Whole-Cell Biosensor with Higher Sensitivity toward Genotoxic Compounds than Constructs Based on recA, umuDC, or sulA Promoters

Anders Norman, Lars Hestbjerg Hansen,* and Søren J. Sørensen

Department of Microbiology, University of Copenhagen, 1307 Copenhagen K, Denmark

Received 19 October 2004/ Accepted 30 November 2004

Four different green fluorescent protein (GFP)-based whole-cell biosensors were created based on the DNA damage inducible SOS response of Escherichia coli in order to evaluate the sensitivity of individual SOS promoters toward genotoxic substances. Treatment with the known carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) revealed that the promoter for the ColD plasmid-borne cda gene had responses 12, 5, and 3 times greater than the recA, sulA, and umuDC promoters, respectively, and also considerably higher sensitivity. Furthermore, we showed that when the SOS-GFP construct was introduced into an E. coli host deficient in the tolC gene, the minimal detection limits toward mitomycin C, MNNG, nalidixic acid, and formaldehyde were lowered to 9.1 nM, 0.16 µM, 1.1 µM, and 141 µM, respectively, which were two to six times lower than those in the wild-type strain. This study thus presents a new SOS-GFP whole-cell biosensor which is not only able to detect minute levels of genotoxins but, due to its use of the green fluorescent protein, also a reporter system which should be applicable in high-throughput screening assays as well as a wide variety of in situ detection studies.

* Corresponding author. Mailing address: Department of Microbiology, University of Copenhagen, Sølvgade 83H, 1307 Copenhagen K, Denmark. Phone: 45 35 32 20 53. Fax: 45 35 32 20 40. E-mail: hestbjerg{at}bi.ku.dk.

Applied and Environmental Microbiology, May 2005, p. 2338-2346, Vol. 71, No. 5
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.5.2338-2346.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gillor, O., Vriezen, J. A. C., Riley, M. A. (2008). The role of SOS boxes in enteric bacteriocin regulation. Microbiology 154: 1783-1792 [Abstract] [Full Text]  
  • Cascales, E., Buchanan, S. K., Duche, D., Kleanthous, C., Lloubes, R., Postle, K., Riley, M., Slatin, S., Cavard, D. (2007). Colicin Biology. Microbiol. Mol. Biol. Rev. 71: 158-229 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»